Bio-Rad Bio-Plex Pro™ Human Th17 Cytokine Assays - Manual (Page 22)

20
Tables 9–10 summarize the volumes required to prepare 1x detection
antibodies from a single 20x stock. Also shown are volumes to prepare 1x
antibodies when mixing two 20x stocks from singleplex assays.
Preparing 1x detection antibodies from 20x stock
Table 9. Premixed panel or one singleplex assay.
Table 10. Mixing singleplex assays.
5. After incubating the beads, samples, standards, blank, and controls,
slowly remove and discard the sealing tape.
6. Wash the wells three times with 100 µl wash buffer according to the
wash method of choice.
7. Vortex the diluted (1x) detection antibodies gently for 5 sec. Pour into
a reagent reservoir and transfer 25 µl to each well of the assay plate
using a multichannel pipet.
8. Cover and incubate in the dark for 30 min at RT with shaking at
850 ±50 rpm.
20x Detection Detection Antibody
# of Wells Antibodies, µl Diluent, µl Total Volume, µl
96 145 2,755 2,900
48 73 1,377 1,450
20x Detection 20x Detection Detection
Antibodies, µl Antibodies, µl Antibody
# of Wells Singleplex #1 Singleplex #2 Diluent, µl Total Volume, µl
96 145 145 2,610 2,900
48 73 73 1,304 1,450